QUALITY IMPROVEMENT PARTHENOCARPIC FRUITS (NO SEEDS)

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Kudret Cihan
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1 QUALITY IMPROVEMENT Longer Shelf Life Products: tomato, melon, flowers Dry Matter Improvement: low calorie chips, low cost processing Low or non-calorie Sweeteners: Thaumantin & monellin expression Better Food Quality Products Altered Fatty Acid content in oil seeds (γ-linolenic acid) Altered Amino-acid & Protein Content cereals (low lysine), legumes (low methionine) Lysine content in animal feeds New gluten subunits in wheat (bread making quality)

Products Altered Fatty Acid content in oil seeds (γ-linolenic acid) Altered Amino-acid & Protein Content cereals

2 QUALITY IMPROVEMENT PARTHENOCARPIC FRUITS (NO SEEDS) Eggplant Tobacco

3 KALİTE İYİLEŞTİRME Methionine S-Adenosyl methionine Transgenik bitkilerde Antisense Antisense teknolojisi ile AAC oksidaz enzimi inhibe edilerek olgunlaşman manın kontol altına alınmas nması. N TR ACS 1-Aminocyclopropane-1 carboxylic acid (ACC) TR Normal ve transgenik kavunlar polinasyondan 38 gün Normal ve transgenik karanfillerin kesildikten sonra hasat edilip 25 derecede 10 gün bekletilmişlerdir. 8 gün sonraki görünümleri. N ACO ETHYLENE Genetik manipülasyonlar ile çiçek şeklinin değiştirlmesi Hüseyin Avni ÖKTEM, Mayıs 1997 N TR

N TR ACS 1-Aminocyclopropane-1 carboxylic acid (ACC) TR Normal ve transgenik kavunlar polinasyondan 38 gün Normal ve transgenik

4 UZUN RAF ÖMRÜ KESİMDEN 8 GÜN SONRA KESİMDEN 24 GÜN SONRA NORMAL TRANSGENİK

5 0 4 hafta 8 hafta N TR UZUN RAF ÖMRÜ

6 ALTIN PİRİNÇP Dünya genelinde 150 milyon çocuk vitamin A eklikliği çekmekte. Güneydoğu Asyada her yıl vitamin A eksikliğine bağlı: - 5 milyon cocuk görme özürlü olmakta - 2 milyon cocuk ölmekte Ana besinleri PİRİNÇ Çözüm: Gen transferi yöntemiyle pirinçte Vitamin A oranının arttırılması kg pirinç tüketimiyle sorun çözümlenebilecek

olmakta - 2 milyon cocuk ölmekte Ana besinleri PİRİNÇ Çözüm: Gen transferi

7 GOLDEN RICE -Worldwide 150 million childiren suffers from Vitamin-A defficiency -South East Asia: 5 million childiren become blined and 2 millon dies. - Main food is rice. - Engineer Vit-A precursor beta-caroten biosynthetic pathways in rice kg/year consumption would solve Vit-A defficiency problem.

8 TR PLANTS AS LIVING BIOREACTORS Industrial Enzymes : α-amylase (liquefaction, bread baking), phytase (animal feeds, replace inorganic P addition) Pharmaceutical Proteins: Antibodies Enkephalins Interferons Glucocerebrosidease (Gaucher s disease therapy) Polymers: Polyhydroxybutyrate (biodegredable polymer) Oral Vaccines Animal Use (Mink enteritis virus, DK, UK, NL, 1997) Human Use (Cholera toxin subunits, USA, 1998) AN APPLE A DAY KEEPS THE DOCTOR AWAY

Glucocerebrosidease (Gaucher s disease therapy) Polymers: Polyhydroxybutyrate (biodegredable polymer) Oral

9 MOLECULAR FARMING Production of pharmaceuitcally important and commercially valuable proteins in plants 3 Main area Antibodies (plantibodies) i. Therapeutic ii. Diagnostic Oral vaccines (edible vaccines)- Human, animal Biopharmaceutiacls for human health

11 PLANT BASED ORAL VACCINES: Cholera Toxin B Subunit Cells are protected by the antiserum Phys. saline Chol. Toxin A B Unimmunized Orraly Immunized Fluid accumulation in the intestine means toxin is effective. No or little fulid accumulation in immunized mice. Means toxin is ineffective % Protection A: Normal vero cell morphology protected by antiserum from orally immunized mice. B: Effect of cholera toxin on vero cells. Effect of oral immunization with bacterial or plant cholera toxin B subunit. Arakawa T., Nat.Biotech. Vol.16, 1998

No or little fulid accumulation in immunized mice.

12 AĞIZDAN ALINABİLECEK AŞILAR Plant-derived vaccine protects target animals against a viral disease Dalsgaard et al., Nature Biotechnology, 1997 Efficacy of a food plant-based oral cholera toxin B subunit vaccine Arakawa et al., Nature Biotechnology, 1998 A plant-derived edible vacine against hepatitis B virus Kapusta et al., FASEB J., 1999 Transgenic plants for production of edible vaccines and antibodies for immunotheraphy Sharma et al., Current Science., 1999 Plants for delivery of edible vaccines. Walmsley, Curr.Opp. Plant Sci., 2000 Development of oral vaccines for human use.. Fooks., Curr.Opp. Mol. Therap., 2000

, Nature Biotechnology, 1998 A plant-derived edible vacine against hepatitis B virus Kapusta et al., FASEB J.

13 YENİLEBİLİR İNSAN ve HAYVAN AŞILARI HEPATİT KOLERA ŞAP (feet and mouth disease) AİDS

14 Foot-and-Mouth Disease Vaccine Production in Transgenic Plants

15 Foot-and-Mouth Disease One of the most contagious animal diseases Permanent impairment of milk production, abortion, high mortality of young cattle and piglets, permanent loss of weight, myocarditis in cattle, pig Important economic losses; e.g. 10 billion in UK in endemic 2001

young cattle and piglets, permanent loss of weight, myocarditis in

16 Foot-and-Mouth Disease Virus FMDV, an Apthovirus; Picornaviridae Nonenveloped, icosohedral capsid SS(+)RNA Serotypes A, O, C, Asia1

17 Usage of TR plants Expression of antigens in TR plants increasingly used for production of immunogens TR plants can be parenterally administered or used as edible vaccines Low concentration of the expressed protein to induce significant immune response Increasing translational efficiency Utilization of plant-optimized synthetic genes via inclusion of specific promoters, enhancers or signal sequences...increased levels of expression of rec genes

induce significant immune response Increasing translational efficiency Utilization of plant-optimized synthetic

18 FMDV vaccine in TR plants First peptide-based vaccine production in TR plants for FDMV Structural protein VP1, carrying critical epitopes responsible for induction of protective neutralizing antibodies,expressed in alfalfa

VP1, carrying critical epitopes responsible for

19 CaMV 35S used as promoter Obtained vector code for rec product VP-betaGUS, fusion protein containing FMDV VP1 followed by gus A gene Introduced into A.tumefaciens by electroporation, alfalfa cocultivated

FMDV VP1 followed by gus A gene Introduced into A.

20 Results 5% of tested individuals presented high beta-gus activity Also showed highest VP1 peptide accumulation Rec VP-betaGUS reached concentrations of mg/g of total

21 Graphics

22 Results Intraperitoneally immunized mice present specific antibody response against both synthetic peptide VP1 and purified FMDV particles VP-betaGUS immunized mice challenged with infectious FMDV, 100% protection were observed

23 In brief Idea of production of vaccines with TR plants cost-friendly and safe Promising results obtained to date, methodolgy to be perfected by using more sophisticated vector systems, appropriate plant Extensive trials to prove effect of FMDV vaccine in meat-producing animals

24 Kanser terapisinde kullanılabilecek antikorların üretimi Transient expression of a tumor specific single chain fragment and a chimeric antibody in tobacco leaves Vaquero et al., PNAS Otoimmün hastalıkların tedavisi Supression of autoimmune diabetes by a plant-delivered cholera toxin B subunithuman glutamate decarboxylase fusion protein. Arakawa et al. Transgenics, 1999

25 Bt Cholesterol oksidaz VI P amilaz inhibitör ü Böcek Direnci Avidin proteaz inhibitörü Raundop Viral PPT Fungal Bakteriyel Hastalık Direnci Sıca k Ağır metal toksisitesi Tuz Abiotik Stres Direnci Makro-element eksikliği Soğuk Kuraklı k Yüksek vitamin içeren gıdalar Kuru madde Miktarının Arttırılması Herbisit Direnci TRANSGENİK BİTKİ TEKNOLOJİSİ Nematod Direnci Gıda Değerinin İyileştirilmesi Domates Ağız yoluyla alınabilen AŞILAR Kanser ve otoimmün hastalıkların tedavisi Uzun Raf Ömrü Endüstriyel Önemi Olan Enzimlerin Üretilmesi Çiçe k Meyve

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